Interaction Details


Details of experiments between Q9P0K7 and P63104
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: RAI14
Fragment: 288-297
Site: 14-3-3_pSer293
Modification: (SEP)293
Sequence: VSSPRSITST
Construct: VSSPR(pSer)ITST (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.06
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 1.99
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 2

Protein A

Protein: RAI14
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSITSTPLS
Construct: PRSIT(pSer)TPLS (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.35
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 4.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.6
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 3

Protein A

Protein: RAI14
Fragment: 295-304
Site: 14-3-3_pSer300
Modification: (SEP)300
Sequence: TSTPLSGKES
Construct: TSTPL(pSer)GKES (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.01
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 0.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 4

Protein A

Protein: RAI14
Fragment: 313-322
Site: 14-3-3_pSer318
Modification: (SEP)318
Sequence: KAEISSIREN
Construct: KAEIS(pSer)IREN (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.05
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 0.74
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 5

Protein A

Protein: RAI14
Fragment: 353-362
Site: 14-3-3_pSer358
Modification: (SEP)358
Sequence: QAKVASLTLH
Construct: QAKVA(pSer)LTLH (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.22
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 3.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 6

Protein A

Protein: RAI14
Fragment: 414-423
Site: 14-3-3_pSer419
Modification: (SEP)419
Sequence: SVLIHSLGKS
Construct: SVLIH(pSer)LGKS (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.02
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 0.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 7

Protein A

Protein: RAI14
Fragment: 662-671
Site: 14-3-3_pSer667
Modification: (SEP)667
Sequence: YRKRKSLEDV
Construct: YRKRK(pSer)LEDV (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.05
Holdup BI standard deviation: 1.35
Immobilized partner concentration (10-6M): 132
P value: 0.02
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 8

Protein A

Protein: RAI14
Fragment: 246-255
Site: 14-3-3_pThr251
Modification: (TPO)251
Sequence: DLKTPTKPKQ
Construct: DLKTP(pThr)KPKQ (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.05
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 1.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 9

Protein A

Protein: RAI14
Fragment: 325-334
Site: 14-3-3_pThr330
Modification: (TPO)330
Sequence: RLSDSTTGAD
Construct: RLSDS(pThr)TGAD (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.14
Holdup BI standard deviation: 0.51
Immobilized partner concentration (10-6M): 132
P value: 0.22
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 10

Protein A

Protein: RAI14
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: GKPGETSPPD
Construct: GKPGE(pThr)SPPD (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 0.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))